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【目的】筛选对百合枯萎病具有抑菌活性的拮抗细菌,对其抑菌活性物质进行初步分离纯化分析。【方法】以强致病力的百合尖孢镰刀菌(Fusarium oxysporum)为靶菌,采用系列稀释法和平板对峙法初筛拮抗细菌,并通过产铁载体能力、水解酶活性、土壤定殖力等多种生防特性指标进行复筛,结合形态学特征、生理生化指标和16S r RNA基因序列比对鉴定其分类地位;利用百合尖孢镰刀菌作为靶菌进行活性追踪,结合酸沉淀、快速柱色谱、HPLC等分离纯化手段,对菌发酵液中的抑菌活性物质进行纯化分析。【结果】在64株百合根际细菌和386株海洋细菌中进行初筛,得到9株对百合镰刀菌具有较强拮抗活性的菌株,最后筛选了1株拮抗活性较强且产铁载体能力和水解酶活性、土壤定殖能力较高的菌株11B91,鉴定为解淀粉芽孢杆菌(Bacillus amyloliquefaciens),其抑菌活性物质初步推测可能为Iturin和Fengycin脂肽类化合物。【结论】菌株11B91在百合枯萎病的生物防治中具有潜在的应用价值,证实海洋来源的微生物也具有防治陆地植物病原菌的潜力,为植物病害的防治拓宽了思路。
【Objective】 The objective of this study was to screen antagonistic bacteria with antibacterial activity against Fusarium wilt and to isolate and purify its antibacterial active substance. 【Method】 With the strong pathogenicity of Fusarium oxysporum as target bacteria, the bacterial strains were screened by serial dilution method and plate confrontation method. Through iron-producing ability, hydrolytic enzyme activity, soil fertility And other biological characteristics of anti-screening index screen, combined with morphological characteristics, physiological and biochemical indicators and 16S rRNA gene sequence alignment to identify its taxonomic status; Fusarium oxysporum Fusarium as a target bacteria activity tracking, combined with acid precipitation, rapid Column chromatography, HPLC and other means of separation and purification, antibacterial active substances in the fermentation broth purification analysis. 【Result】 Nine strains of rhizobacteria were isolated from 64 Lily rhizobacteria and 386 strains of marine bacteria. Nine strains with strong antagonistic activities against Fusarium spermatophore were obtained. One strain with strong antagonistic activity and iron-producing ability Hydrolytic enzyme activity, soil colonization higher strain 11B91, identified as Bacillus amyloliquefaciens (Bacillus amyloliquefaciens), the antibacterial active substance may be presumed to Iturin and Fengycin lipopeptide compounds. 【Conclusion】 Strain 11B91 has potential value in the biological control of Fusarium wilt and confirmed that marine-derived microorganisms have the potential of preventing and controlling terrestrial plant pathogens and broadened the ideas for the prevention and treatment of plant diseases.